Integrated microfluidic system for the processing of tissues into cellular suspensions

Abstract

A microfluidic system for processing a tissue sample includes a microfluidic digestion device having an outlet fluidically connected to an inlet of a dissociation/filter device. The microfluidic digestion device includes an inlet and an outlet and a tissue chamber that connects to plurality of upstream fluidic channels and a plurality of downstream fluidic channels. The microfluidic dissociation/filter device includes an inlet, a first outlet, a second outlet, and a plurality of furcating dissociation channels having a plurality of expansion and constriction regions disposed along a length thereof, wherein one or more filters are disposed in a flow path downstream of the plurality of furcating dissociation channels. Pumps are provided to pump buffer and/or enzyme-containing fluid through the digestion device and dissociation/filter device. Tissue is initially processed in the digestion device and then passes into the dissociation/filter device.

What is claimed is: 
     
         1 . A microfluidic system for processing a tissue sample comprising:
 a microfluidic digestion device comprising an inlet and an outlet and a flow path defined between the inlet and the outlet, the flow path comprising a tissue chamber configured to hold the tissue sample and a plurality of upstream fluidic channels communicating with the tissue chamber on the inlet side of the flow path and a plurality of downstream fluidic channels communicating with the tissue chamber on the outlet side of the flow path;   a first pump configured to pump a buffer-containing fluid and/or an enzyme-containing fluid into the inlet of the microfluidic digestion device;   a microfluidic dissociation/filter device comprising an inlet, a first outlet, a second outlet, and a flow path defined between the inlet and the outlet, the flow path comprising a plurality of furcating dissociation channels having a plurality of expansion and constriction regions disposed along a length thereof, wherein one or more filters are disposed in the flow path downstream of the plurality of furcating dissociation channels;   a second pump configured to pump a buffer-containing or other fluid into the inlet of the microfluidic dissociation/filter device; and   wherein the outlet of the microfluidic digestion device is fluidically coupled to the inlet of the microfluidic dissociation/filter device.   
     
     
         2 . The microfluidic system of  claim 1 , further comprising one or more valves interposed between the outlet of the microfluidic digestion device and the inlet of the microfluidic dissociation/filter device. 
     
     
         3 . The microfluidic system of  claim 1 , wherein the first outlet comprises a valve or cap to close the same and wherein when the first outlet of the microfluidic dissociation/filter device is closed fluid causes fluid to flow through a flow path containing the one or more filters. 
     
     
         4 . The microfluidic system of  claim 1 , wherein the second outlet comprises a valve or cap to close the same and wherein when the second outlet of the microfluidic dissociation/filter device is closed causes fluid to exit the microfluidic dissociation/filter device via the first outlet without passing through the one or more filters. 
     
     
         5 . The microfluidic system of  claim 1 , wherein the one or more filters comprises a first filter having a pore size within the range of about 50-100 μm and a second filter having a pore size within the range of about 15-50 μm. 
     
     
         6 . The microfluidic system of  claim 1 , wherein the number of upstream fluidic channels equals the number of downstream fluidic channels. 
     
     
         7 . The microfluidic system of  claim 6 , wherein the upstream and downstream fluidic channels have a width within the range between about 250 μm and 750 μm. 
     
     
         8 . The microfluidic system of  claim 1 , further comprising a port disposed on the microfluidic digestion device and in communication with the tissue chamber. 
     
     
         9 . A method of using the microfluidic system of  claim 1  comprising:
 loading the tissue sample into the tissue chamber of the microfluidic digestion device; 
 pumping the buffer-containing fluid and/or an enzyme-containing fluid into the inlet of the microfluidic digestion device with the first pump; 
 transferring fluid containing processed tissue sample to the microfluidic dissociation/filter device; 
 pumping a buffer-containing or other fluid into the inlet of the microfluidic dissociation/filter device along with the processed tissue from the microfluidic digestion device with the second pump; and 
 collecting effluent from the second outlet of the microfluidic dissociation/filter device. 
 
     
     
         10 . The method of  claim 9 , wherein pumping the buffer-containing fluid and/or an enzyme-containing fluid into the inlet of the microfluidic digestion device with the first pump comprises recirculating fluid into the microfluidic digestion device with the first pump. 
     
     
         11 . The method of  claim 9 , wherein the enzyme-containing fluid comprises a fluid containing collagenase. 
     
     
         12 . The method of  claim 9 , wherein the pumping the buffer-containing fluid and/or an enzyme-containing fluid into the inlet of the microfluidic digestion device is performed in intervals where effluent is removed from the microfluidic digestion device at the end of each interval and replacement enzyme-containing fluid is pumped into the microfluidic digestion device. 
     
     
         13 . The method of  claim 9 , wherein total processing time in the microfluidic digestion device and the microfluidic dissociation/filter device is 1 minute or more. 
     
     
         14 . The method of  claim 9 , wherein total processing time in the microfluidic digestion device and the microfluidic dissociation/filter device is 15 minutes or more. 
     
     
         15 . The method of  claim 9 , wherein the processed tissue from the microfluidic digestion device is recirculated through the plurality of furcating dissociation channels in the microfluidic dissociation/filter device a plurality of times prior to exit from the second outlet. 
     
     
         16 . The method of  claim 9 , wherein the microfluidic dissociation/filter device contains a single filter. 
     
     
         17 . The method of  claim 9 , wherein the microfluidic dissociation/filter device contains a plurality of filters.